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RIPK4 Downregulation Reduces ABCG2 Expression, Increasing BRAF-Mutated Melanoma Cell Susceptibility to Cisplatin- and

Bartlomiej Olajossy1,2, Norbert Wronski1,2, Ewelina Madej1

  • 1Department of Biophysics and Cancer Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa Street 7, 30-387 Krakow, Poland.

Biomolecules
|January 8, 2025
PubMed

Insights

Receptor-Interacting Protein Kinase 4 (RIPK4) influences melanoma chemoresistance. Downregulating RIPK4 enhances sensitivity to cisplatin and doxorubicin, suggesting RIPK4

Area of Science:

  • Oncology
  • Molecular Biology
  • Cancer Research

Background:

  • Melanoma exhibits resistance to standard chemotherapies like cisplatin (CisPt) and doxorubicin (DOX).
  • Aberrant Receptor-Interacting Protein Kinase 4 (RIPK4) expression is linked to melanoma growth via NFκB and Wnt/β-catenin pathways, which influence chemoresistance.
  • The specific role of RIPK4 in melanoma response to chemotherapeutics remains uninvestigated.

Purpose of the Study:

  • To investigate the impact of RIPK4 modulation (downregulation and overexpression) on the sensitivity of BRAF-mutated melanoma cells to CisPt and DOX.
  • To elucidate the underlying molecular mechanisms governing RIPK4's role in melanoma chemoresistance.

Main Methods:

  • Utilized siRNA and CRISPR/Cas9 for RIPK4 silencing and dCas9-VPR for overexpression in A375 and WM266.4 melanoma cell lines.
  • Assessed CisPt and DOX-induced apoptosis via caspase 3/7 activity, annexin V/7AAD staining, and flow cytometry (FASC).
  • Employed qRT-PCR and Western blotting to analyze apoptosis-related genes/proteins (cleaved PARP, p53, cyclin D1).

Main Results:

  • RIPK4 overexpression inhibited, whereas RIPK4 downregulation significantly enhanced, apoptosis induced by CisPt or DOX in melanoma cells.
  • RIPK4 downregulation effects mimicked those of cyclosporin A, an ABCG2 inhibitor, suggesting a link to drug efflux mechanisms.
  • Preliminary evidence indicated crosstalk between RIPK4, BIRC3, and ABCG2, potentially mediating chemoresistance.

Conclusions:

  • RIPK4 plays a crucial role in regulating chemoresistance in BRAF-mutated melanoma cells treated with cisplatin and doxorubicin.
  • Targeting RIPK4, particularly through its downregulation, represents a potential strategy to overcome chemoresistance in melanoma.
  • Further research into the RIPK4, BIRC3, and ABCG2 interplay is warranted to fully understand and exploit this pathway for therapeutic benefit.
Keywords:
MPRRIPK4cisplatincyclosporin Adoxorubicinmelanoma

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